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枯草芽孢杆菌甲基丙二酸半醛脱氢酶的表达、纯化、结晶及初步X射线衍射数据

Expression, purification, crystallization and preliminary X-ray diffraction data of methylmalonate-semialdehyde dehydrogenase from Bacillus subtilis.

作者信息

Dubourg Hélène, Stines-Chaumeil Claire, Didierjean Claude, Talfournier François, Rahuel-Clermont Sophie, Branlant Guy, Aubry André

机构信息

Laboratoire de Cristallographie et Modélisation des Matériaux Minéraux et Biologiques, UMR 7036 CNRS-UHP, Groupe Biocristallographie, Faculté des Sciences et Techniques, BP 239, 54506 Vandoeuvre-lès-Nancy, France.

出版信息

Acta Crystallogr D Biol Crystallogr. 2004 Aug;60(Pt 8):1435-7. doi: 10.1107/S0907444904012533. Epub 2004 Jul 21.

Abstract

Methylmalonate-semialdehyde dehydrogenase from Bacillus subtilis was cloned and overexpressed in Escherichia coli. Suitable crystals for X-ray diffraction experiments were obtained by the hanging-drop vapour-diffusion method using ammonium sulfate as precipitant. The crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 195.2, b = 192.5, c = 83.5 A, and contain one tetramer per asymmetric unit. X-ray diffraction data were collected to 2.5 A resolution using a synchrotron-radiation source. The crystal structure was solved by the molecular-replacement method.

摘要

来自枯草芽孢杆菌的甲基丙二酸半醛脱氢酶在大肠杆菌中被克隆并过量表达。通过使用硫酸铵作为沉淀剂的悬滴气相扩散法获得了适合X射线衍射实验的晶体。这些晶体属于空间群P2(1)2(1)2(1),晶胞参数a = 195.2、b = 192.5、c = 83.5 Å,每个不对称单元包含一个四聚体。使用同步辐射源收集了分辨率为2.5 Å的X射线衍射数据。通过分子置换法解析了晶体结构。

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